CN105388200A - Method for preparing sensor used for organic phosphorus pesticide detection - Google Patents
Method for preparing sensor used for organic phosphorus pesticide detection Download PDFInfo
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- CN105388200A CN105388200A CN201510667756.6A CN201510667756A CN105388200A CN 105388200 A CN105388200 A CN 105388200A CN 201510667756 A CN201510667756 A CN 201510667756A CN 105388200 A CN105388200 A CN 105388200A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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- G01N27/42—Measuring deposition or liberation of materials from an electrolyte; Coulometry, i.e. measuring coulomb-equivalent of material in an electrolyte
- G01N27/423—Coulometry
Abstract
The invention provides a method for preparing an enzyme sensor used for organic phosphorus pesticide detection. The sensor comprises a biological enzyme layer, a nano particle/conductive polymer layer and an electrode substrate. Nano particles are Au-Fe3O4. A conductive polymer is a quinines sulphonate/graphene oxide composite material. The electrode substrate is made of a glass carbon material. An enzyme solution is an acetycholinesterase and chitosan-acetic acid solution. The linear detection range of the prepared sensor on existing organic phosphorus pesticide is from 4.0*10-13 mol/L to 5.0*10-9 mol/L. The detection limit of the sensor on the existing organic phosphorus pesticide is 1.0*10-13 mol/L.
Description
Technical field
The present invention relates to a kind of chemical sensor, specifically a kind of preparation method of the sensor for detection of organic phosphorus pesticide.
Background technology
Organophosphorus pesticide mainly refers to phosphate, phosphorothionate, phosphoramidate organophosphorus compounds, because its insecticide efficiency is high, little to the poisoning of plant, the feature such as persistence is low in the environment, become the class agricultural chemicals that China is the most frequently used at present, be widely used in various crops especially vegetables and fruits.But remains of pesticide enters in body along with food chain, can suppress the activity of cholinesterase in human body, causes the metabolic disorder of nerve conduction medium, cause all kinds of acute, slow poisoning situation, as ataxia, stupor, paralysis are even dead.Therefore timely, accurate, sensitive monitoring and detection are carried out to organophosphorus pesticide, become pressing issues urgently to be resolved hurrily.
Traditional pesticide residue detection method mainly contains: GC-MS(gas chromatography-mass spectrography), high performance liquid chromatography, spectroscopic methodology, immunoassay, chemoluminescence method etc.Although the residual quantity of agricultural chemicals can be detected more exactly, there is apparatus expensive, the shortcomings such as sample-pretreating method is loaded down with trivial details, analytical cycle is long.
Summary of the invention
The object of the invention is for prior art above shortcomings, a kind of sensor of quick, sensitive, easy detection of organic phosphorus pesticide is provided.This sensor be based on the high specificity of bio-identification and nano material to the amplification of electrochemical detection signal, biological chemistry, nano material, sensing technology three are combined, construct the enzyme sensor that Novel composite nano material is modified.
The present invention is achieved by the following technical solutions, and the enzyme sensor that the present invention relates to for detection of organic phosphorus pesticide comprises: biological enzyme layer, nano particle conductive polymer coating and electrode matrix.
For a preparation method for the enzyme sensor of detection of organic phosphorus pesticide, it is characterized in that, comprise the following steps:
The first step: Au-Fe
3o
4the preparation of nano particle:
Get Fe
3o
4powder ultrasonic is scattered in redistilled water, adds sodium citrate solution, slowly adds oxammonium hydrochloride and gold chloride while stirring, stirs certain hour, namely obtains Au-Fe
3o
4the dispersion soln of composite nanoparticle;
Second step: Au-Fe
3o
4the preparation of the acetylcholine enzyme biologic sensor of nano particle/conducting polymer modified:
First glass-carbon electrode is carried out ultrasonic cleaning in redistilled water after polishing; Then ultrasonic cleaning in acetone, sodium hydroxide solution, nitric acid and redistilled water successively;
Then the glass-carbon electrode after cleaning puts into M polymer monomer, the water-soluble quinones sulfonate of M and graphene oxide solution, logical nitrogen is after 30 minutes, three-electrode system is adopted to carry out continuous current electro-deposition, then gained modified electrode is carried out electrochemical reduction, obtain conducting polymer/quinones sulfonated bodies/graphene oxide composite material modified electrode;
Get a certain amount of Au-Fe
3o
4nanoparticle dispersion solution drips the conducting polymer/quinones sulfonated bodies/graphene oxide composite material modified electrode surface being coated in and drying, naturally dry, AChE is mixed with isopyknic chitosan-acetic acid solution, and get mixed solution 1 μ L-20 μ L and drip and be coated on modified electrode, naturally dry, obtain Au-Fe
3o
4the acetylcholine enzyme biologic sensor of nano particle/conducting polymer modified.
Described quinones sulfonate is anthraquinone or phenanthrenequione.
Described M polymer monomer is the one in pyrroles, aniline, 3,4-rthylene dioxythiophene.Described redistilled water refers to: the pure water obtained after twice distillation.
Described sodium citrate solution refers to: solution concentration is 0.05mol/L-1mol/L.
Described oxammonium hydrochloride refers to: solution concentration is 0.05mol/L-1mol/L.
Described gold chloride refers to: mass concentration is 0.5%-10%.
Described stirring certain hour refers to: 5-30 hour.
Get 2 μ L-20 μ LAu-Fe
3o
4nanoparticle dispersion solution drips the conducting polymer/quinones sulfonated bodies/graphene oxide composite material modified electrode surface being coated in and drying, naturally dry, the AChE (100U/mL) of 1 μ L-20 μ L is mixed with isopyknic 0.5mg/mL-5mg/mL chitosan-acetic acid solution, and get mixed solution 1 μ L-20 μ L and drip and be coated on modified electrode, naturally dry, obtain Au-Fe
3o
4the acetylcholine enzyme biologic sensor of nano particle/conducting polymer modified.
Described quinones sulfonate is the one of anthraquinone or phenanthrenequione.
Described M polymer monomer is the one in pyrroles, aniline, 3,4-rthylene dioxythiophene.
The present invention is detected by following steps:
Acetylcholine ester substrate for enzymatic activity acetylthiocholine (ATCh) is hydrolyzed, and generate product thiocholine and acetic acid, thiocholine is easily oxidized under specific potential, generation current, current signal size and thiocholine concentration linear.
CHI832 type electro-chemical systems is utilized to record AChE/Au-SiO under+0.6V
2/ GCE sensor is to the i-t response of variable concentrations ATCh and working curve;
Under+0.6V operating potential, after sensor signal is stablized, in PBS, add the ATCh of concentration known, stir 5.0min, after current stabilization, in static solution, record produced steady-state current I
1; In the ATCh solution of another part of similarity condition, add certain density agricultural chemicals, after leaving standstill a few minutes after stirring, measure the steady-state current I that there is various variable concentrations organophosphorus pesticide
2.Activity inhibited due to enzyme causes electric current to decline, and the degree that electric current declines is directly proportional to the concentration of the organophosphorus pesticide in solution, and inhibiting rate (I
1-I
2)/I
1linear with the logarithm of organic phosphorus concentration, the concentration of organophosphorus pesticide therefore can be detected by the reduction measuring enzymatic activity.
The method of the invention can realize the detection of organophosphorus pesticide concentration with easy steps, the detection range of linearity of this sensor to existing machine phosphorus insecticide is 4.0 × 10
-13-5.0 × 10
-9mol/L, the detection of sensor to existing machine phosphorus insecticide is limited to 1.0 × 10
-13mol/L.
Embodiment
Embodiment is implemented under premised on technical solution of the present invention, give detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Embodiment 1:
Get the Fe that 5mg mean diameter is 80nm
3o
4powder ultrasonic is scattered in 50mL redistilled water, adds 4mL0.5mol/L sodium citrate solution, stirs 10min.Slowly add 0.8mL oxammonium hydrochloride (NH while stirring
2oHHCl) (0.3mol/L) and 0.5mL gold chloride (HAuCl
4) (8%), stir 25 hours, namely obtain the Fe of gold doping
3o
4the dispersion soln of composite nanoparticle.
By the Al of naked glass-carbon electrode with 0.5 μm of diameter
2o
3sticking with paste sanding and polishing on chamois leather, make into minute surface, is the HNO of 1:1 in acetone, volume ratio successively
3, ultrasonic process in NaOH (1mol/L) and intermediate water.Then electrode is put into the pyrrole monomer containing 0.1M, 0.0005M phenanthrenequione sulfonate, 0.5mg/ml graphene oxide water solution, logical N
2after 30min, adopt three-electrode system to carry out continuous current electro-deposition, applying electric current is 0.5mAcm
-2, the electricity of electro-deposition is 1.2 coulombs; Carry out electrochemical reduction being placed in three-electrode system after being cleaned by obtained polypyrrole film, applying voltage is 1.1V, and electrolytic solution is PBS buffer solution, repeatedly cleans can obtain conducting polymer modified electrode after reaction with water.Get 8 μ LAu-Fe
3o
4nanoparticle dispersion solution drips the modified electrode surface being coated in and drying, naturally dry, 2 μ LAChE (100U/mL) are mixed with shitosan (CHIT) acetum of isopyknic 1mg/mL, and get 8 μ L mixed solutions and drip on the electrode that is coated in and modified nano particle/conducting polymer, naturally dry, obtain detection of organic phosphorus pesticide sensor.
Embodiment 2:
Get the Fe that 5mg mean diameter is 200nm
3o
4powder ultrasonic is scattered in 50mL redistilled water, adds 4mL1mol/L sodium citrate solution, stirs 10min.Slowly add 0.8mL oxammonium hydrochloride (NH while stirring
2oHHCl) (0.8mol/L) and 0.5mL gold chloride (HAuCl
4) (10%), stir 28 hours, namely obtain the Fe of gold doping
3o
4the dispersion soln of composite nanoparticle.
By the Al of naked glass-carbon electrode with 0.5 μm of diameter
2o
3sticking with paste sanding and polishing on chamois leather, make into minute surface, is the HNO of 1:1 in acetone, volume ratio successively
3, ultrasonic process in NaOH (1mol/L) and intermediate water.Then electrode is put into pyrrole monomer, 0.0005M anthraquinone sulphonates, the 1.0mg/ml graphene oxide water solution containing 0.2M, logical N
2after 30min, adopt three-electrode system to carry out continuous current electro-deposition, applying electric current is 2mAcm
-2, the electricity of electro-deposition is 2.0 coulombs; Carry out electrochemical reduction being placed in three-electrode system after being cleaned by obtained polypyrrole film, applying voltage is 1.2V, and electrolytic solution is PBS buffer solution, repeatedly cleans can obtain conducting polymer modified electrode after reaction with water.Get 13 μ LAu-Fe
3o
4nanoparticle dispersion solution drips the modified electrode surface being coated in and drying, naturally dry, 10 μ LAChE (100U/mL) are mixed with shitosan (CHIT) acetum of isopyknic 1mg/mL, and get 10 μ L mixed solutions and drip on the electrode that is coated in and modified nano particle/conducting polymer, naturally dry, obtain detection of organic phosphorus pesticide sensor.
Embodiment 3:
Get the Fe that 3mg mean diameter is 20nm
3o
4powder ultrasonic is scattered in 35mL redistilled water, adds 1.88mL0.1mol/L sodium citrate solution, stirs 10min.Slowly add 0.07mL oxammonium hydrochloride (NH while stirring
2oHHCl) (1mol/L) and 0.125mL gold chloride (HAuCl
4) (10%), stir 5 hours, namely obtain the Fe of gold doping
3o
4the dispersion soln of composite nanoparticle.
By the Al of naked glass-carbon electrode with 0.5 μm of diameter
2o
3sticking with paste sanding and polishing on chamois leather, make into minute surface, is the HNO of 1:1 in acetone, volume ratio successively
3, ultrasonic process in NaOH (1mol/L) and intermediate water.Then electrode is put into containing aniline monomer, 0.1M phenanthrenequione sulfonate, the 1.0mg/ml graphene oxide water solution containing 0.05M, logical N
2after 30min, adopt three-electrode system to carry out continuous current electro-deposition, applying electric current is 1mAcm
-2, the electricity of electro-deposition is 1.6 coulombs; Carry out electrochemical reduction being placed in three-electrode system after being cleaned by obtained polyaniline film, applying voltage is 1.0V, and electrolytic solution is PBS buffer solution, repeatedly cleans can obtain conducting polymer modified electrode after reaction with water.Get 5 μ LAu-Fe
3o
4nanoparticle dispersion solution drips the naked glassy carbon electrode surface being coated in and drying, naturally dry, 2 μ LAChE (100U/mL) are mixed with shitosan (CHIT) acetum of isopyknic 1mg/mL, and get 2 μ L mixed solutions and drip on the electrode that is coated in and modified nano particle/conducting polymer, naturally dry, obtain detection of organic phosphorus pesticide sensor.
Embodiment 4:
Get the Fe that 3mg mean diameter is 30nm
3o
4powder ultrasonic is scattered in 35mL redistilled water, adds 1.88mL0.1mol/L sodium citrate solution, stirs 10min.Slowly add 0.07mL oxammonium hydrochloride (NH while stirring
2oHHCl) (0.1mol/L) and 0.125mL gold chloride (HAuCl
4) (1%), stir 20 hours, namely obtain the Fe of gold doping
3o
4the dispersion soln of composite nanoparticle.
By the Al of naked glass-carbon electrode with 0.5 μm of diameter
2o
3sticking with paste sanding and polishing on chamois leather, make into minute surface, is the HNO of 1:1 in acetone, volume ratio successively
3, ultrasonic process in NaOH (1mol/L) and intermediate water.Then electrode is put into pyrrole monomer, 0.0005M anthraquinone sulphonates, the 1.0mg/ml graphene oxide water solution containing 0.2M, logical N
2after 30min, adopt three-electrode system to carry out continuous current electro-deposition, applying electric current is 2mAcm
-2, the electricity of electro-deposition is 2.0 coulombs; Carry out electrochemical reduction being placed in three-electrode system after being cleaned by obtained polypyrrole film, applying voltage is 1.2V, and electrolytic solution is PBS buffer solution, repeatedly cleans can obtain conducting polymer modified electrode after reaction with water.Get 5 μ LAu-Fe
3o
4nanoparticle dispersion solution drips the naked glassy carbon electrode surface being coated in and drying, naturally dry, 2 μ LAChE (100U/mL) are mixed with shitosan (CHIT) acetum of isopyknic 1mg/mL, and get 2 μ L mixed solutions and drip on the electrode that is coated in and modified nano particle/conducting polymer, naturally dry, obtain detection of organic phosphorus pesticide sensor.
Claims (3)
1. for a preparation method for the enzyme sensor of detection of organic phosphorus pesticide, it is characterized in that, comprise the following steps:
The first step: Au-Fe
3o
4the preparation of nano particle:
Get Fe
3o
4powder ultrasonic is scattered in redistilled water, adds sodium citrate solution, slowly adds oxammonium hydrochloride and gold chloride while stirring, stirs certain hour, namely obtains Au-Fe
3o
4the dispersion soln of composite nanoparticle;
Second step: Au-Fe
3o
4the preparation of the acetylcholine enzyme biologic sensor of nano particle/conducting polymer modified:
First glass-carbon electrode is carried out ultrasonic cleaning in redistilled water after polishing; Then ultrasonic cleaning in acetone, sodium hydroxide solution, nitric acid and redistilled water successively;
Then the glass-carbon electrode after cleaning puts into M polymer monomer, the water-soluble quinones sulfonate of M and graphene oxide solution, logical nitrogen is after 30 minutes, three-electrode system is adopted to carry out continuous current electro-deposition, then gained modified electrode is carried out electrochemical reduction, obtain conducting polymer/quinones sulfonated bodies/graphene oxide composite material modified electrode;
Get a certain amount of Au-Fe
3o
4nanoparticle dispersion solution drips the conducting polymer/quinones sulfonated bodies/graphene oxide composite material modified electrode surface being coated in and drying, naturally dry, AChE is mixed with isopyknic chitosan-acetic acid solution, and get mixed solution 1 μ L-20 μ L and drip and be coated on modified electrode, naturally dry, obtain Au-Fe
3o
4the acetylcholine enzyme biologic sensor of nano particle/conducting polymer modified.
2. the preparation method of a kind of enzyme sensor for detection of organic phosphorus pesticide according to claim 1, it is characterized in that, described quinones sulfonate is anthraquinone or phenanthrenequione.
3. the preparation method of a kind of enzyme sensor for detection of organic phosphorus pesticide according to claim 1, it is characterized in that, described M polymer monomer is the one in pyrroles, aniline, 3,4-rthylene dioxythiophene.
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| CN201510667756.6A CN105388200B (en) | 2015-10-16 | 2015-10-16 | A kind of transducer production method for detection of organic phosphorus pesticide |
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| CN105388200B CN105388200B (en) | 2018-02-09 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106404756A (en) * | 2016-09-05 | 2017-02-15 | 济南大学 | Preparation method and application of electrochemiluminescence sensor based on graphene/Fe3O4@Au/CeO2/TiO2 |
| CN109324099A (en) * | 2018-08-31 | 2019-02-12 | 盐城工学院 | A kind of volt-ampere of type ammonium ion electrochemica biological sensor and its preparation method and application |
| CN110596218A (en) * | 2019-10-09 | 2019-12-20 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of enzyme sensor for detecting organophosphorus pesticide, product and application thereof |
| CN110632126A (en) * | 2019-09-04 | 2019-12-31 | 东华大学 | Organophosphorus compound sensor and preparation method thereof |
| CN111521658A (en) * | 2020-06-12 | 2020-08-11 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of high-sensitivity sensor for pesticide detection, product and application thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106404756A (en) * | 2016-09-05 | 2017-02-15 | 济南大学 | Preparation method and application of electrochemiluminescence sensor based on graphene/Fe3O4@Au/CeO2/TiO2 |
| CN109324099A (en) * | 2018-08-31 | 2019-02-12 | 盐城工学院 | A kind of volt-ampere of type ammonium ion electrochemica biological sensor and its preparation method and application |
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| CN110596218A (en) * | 2019-10-09 | 2019-12-20 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of enzyme sensor for detecting organophosphorus pesticide, product and application thereof |
| CN111521658A (en) * | 2020-06-12 | 2020-08-11 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of high-sensitivity sensor for pesticide detection, product and application thereof |
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